Complex wounds in dogs are a recurrent problem in veterinary clinical application and can compromise skin healing; in this sense, tissue bioengineering focused on regenerative medicine can be a great ally. Decellularized and recellularized skin scaffolds are produced to be applied in different and complex canine dermal wounds in the present investigation. Dog skin fragments are immersed in a 0.5% sodium dodecyl sulfate (SDS) solution at room temperature and overnight at 4 °C for 12 days. Decellularized samples are evaluated by histological analysis, scanning electron microscopy (SEM) and gDNA quantification. Some fragments are also recellularized using mesenchymal stem cells (MSCs). Eight adult dogs are divided into three groups for the application of the decellularized (Group I, n = 3) and recellularized scaffolds (Group II, n = 3) on injured areas, and a control group (Group III, n = 2). Wounds are evaluated and measured during healing, and comparisons among the three groups are described. In 30- and 60-day post-grafting, the histopathological analysis of patients from Groups I and II shows similar patterns, tissue architecture preservation, epithelial hyperplasia, hyperkeratosis, edema, and mononuclear inflammatory infiltrate. Perfect integration between scaffolds and wounds, without rejection or contamination, are observed in both treated groups. According to these results, decellularized skin grafts may constitute a potential innovative and functional tool to be adopted as a promising dog cutaneous wound treatment. This is the first study that applies decellularized and recellularized biological skin grafts to improve the healing process in several complex wounds in dogs, demonstrating great potential for regenerative veterinary medicine progress.